Molecular Dissection of Functional Structures in the Ribosome
| Project/Area Number |
14035222
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | Niigata University (2003-2006)
Shinshu University (2002) |
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Principal Investigator |
UCHIUMI Toshio Niigata University, Institute of Science and Technology, Professor (50143764)
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| Project Period (FY) |
2002 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥62,600,000 (Direct Cost: ¥62,600,000)
Fiscal Year 2006: ¥12,400,000 (Direct Cost: ¥12,400,000)
Fiscal Year 2005: ¥12,400,000 (Direct Cost: ¥12,400,000)
Fiscal Year 2004: ¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 2003: ¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 2002: ¥12,600,000 (Direct Cost: ¥12,600,000)
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| Keywords | Ribosome / GTPase / Ribosomal protein / rRNA / Translation / Elongation factor / RNA-protein interaction / Archaebacteria / RNA-タンバク質相互作用 / GTPase / IRES / 翻訳 / 蚕 / リボソームRNA / リボソームPタンパク質 / L7 / L12 / L10 |
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| Research Abstract |
The ribosomal GTPase-associated center is composed of a part of 23S rRNA and a few proteins, which assemble onto the rRNA region and form a characteristic complex, termed the stalk. This rRNA-protein complex plays a crucial role in ribosomal interaction with GTP binding translation factors, GTP hydrolysis, and regulation of translation rate. In this project, I analyzed the complex structure by biochemical approaches, and obtained the following new information, particularly on the stalk complex.
1) Establishment of a functional assay system, "hybrid ribosome"
We established conditions for in vitro reconstitution of the stalk complex in Escherichia coli ribosomes and its replacement with its counterparts from other species. This provided a useful system to investigate functional structure of the stalk complex.
2) Assembly mode of eukaryotic PO. P1-P2 stalk complex and its functional characterization
We demonstrated that eukaryotic proteins P1 and P2 form a heterodimer and two dimes bind to the C-terminal region of P0 to form pentameric complex. We also clarified that the stalk complex modulates functional structure of 23S rRNA in the GTPase-associated center.
3) Structural and functional characterization of the archaebacterial stalk complex
We demonstrated that archaebacterial L12 protein forms a homodimer and three dimers bind to the C-terminal region of the anchor protein P0, and that the resultant heptameric complex shows accessibility to eukaryotic as well as archaebacterial translation factors.
4) Structural and functional characterization of the eubacterial stalk complex
We showed that two E. coli L12 homodimers bind to the C-terminal region of the anchor protein L10, whereas three L12 dimers bind to L10 in case of thermophilic eubacteria. We also clarified that a complex variant composed of L10 and one L12 dimer is unstable on the ribosome. This suggests a relationship between number of L12 dimer and stability of the stalk complex in the ribosome.
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Report
(6 results)
Research Products
(171 results)
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[Journal Article] Three binding sites for stalk protein dimers are generally present in ribosomes from archaeal organism.2007
Author(s)
Maki, Y., Hashimoto, T., Zhou, M., Naganuma, T., Ohta, J., Nomura, T., Robinson, C. V., Uchiumi, T.
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Journal Title
J Biol Chem 282
Pages: 32827-32833
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] In vitro reconstitution of the GTPase-associated center of the archaebacterial ribosome: the functional features observed in a hybrid form with Escherichia coli 50S subunits.2006
Author(s)
Nomura, T., Nakano, K., Maki, Y., Naganuma, T., Nakashima, T., Tanaka, I., Kimura, M., Akira Hachimori, A., Uchiumi, T.
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Journal Title
Biochemical J. 396
Pages: 565-571
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] A mode of assembly of PO, P1 and P2 proteins at the GTPaseassociated center in animal ribosome : In vitro analyses with PO truncation mutants.2005
Author(s)
Hagiya, A., Naganuma, T., Maki, Y., Ohta, J., Tohkairin, Y., Shimizu, T., Nomura, T., Hachimori, A., Uchiumi, T.
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Journal Title
J. Biol. Chem 280
Pages: 39193-39199
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Baculovirus-mediated expression and isolation of human ribosomal phosphoprotein P0 carrying a GST-tag in a functional state.2004
Author(s)
Abo, Y., Akiko Hagiya, A., Naganuma, T., Tohkairin, Y., Shiomi, K., Kajiura, Z., Hachimori, A., Uchiumi, T., Nakagaki, M.
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Journal Title
Biochem. Biophys. Res. Commun. 322
Pages: 814-819
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] A point mutation in ribosomal protein L7/L12 reduces its ability to form a compact dimer structure and to assemble into the GTPase center.2003
Author(s)
Nomura, T., Mochizuki, R., Dabbs, E. R., Yoshihiro Shimizu, Y., Ueda, T., Hachimori, A., Uchiumi, T.
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Journal Title
Biochemistry 42
Pages: 4691-4698
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Structural elements in the internal ribosome entry site of Plautia stali intestine virus responsible for binding with ribosomes.2003
Author(s)
Nishiyama, T., Yamamoto, H., Shibuya, N., Hatakeyama, Y., Hachimori, A., Uchiumi, T., Nakashima, N.
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Journal Title
Nucleic Acids Res. 31
Pages: 2434-2442
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Cellular and subcellular distributions of translation initiation, elongation and release factors in rat hippocampus.2003
Author(s)
Inamura, N., Hoshino, S., Uchiumi, T., Nawa, H., Takei, N.
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Journal Title
Brain Res. Mol. Brain Res. 111
Pages: 165-174
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] RALyase ; a terminator of elongation function of depurinated ribosomes.2003
Author(s)
Ozawa, A., Sawasaki, T., Takai, K., Uchiumi, T., Hori, H., Endo, Y.
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Journal Title
FEBS Lett. 555
Pages: 455-458
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Interaction among silkworm ribosomal proteins P1, P2 and PO required for functional protein binding to the GTPase-associated domain of 28S rRNA.2002
Author(s)
Shimizu, T., Nakagaki, M., Nishi, Y., Kobayashi, Y., Hachimori, A., Uchiumi, T.
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Journal Title
Nucleic Acids Res. 30
Pages: 2620-2627
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Autoantibody against ribosomal protein L14 in patients with systemic lupus erythematosus.2002
Author(s)
Hasegawa, H., Uchiumi, T., Sato, T., Saito, A., Nakano, M., Gejyo F.
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Journal Title
Clin. Exp. RheumatoL 20
Pages: 139-144
Description
「研究成果報告書概要(欧文)」より
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